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The opportunistic pathogen Legionella is the leading cause of reported waterborne disease outbreaks in the United States. Legionella can thrive under the warm, stagnant, low-disinfectant conditions characteristic of premise (i.e., building) plumbing systems, making it challenging to identify effective interventions for its control. Copper (Cu) is a promising antimicrobial that can be dosed directly to water via copper-silver ionization systems or released naturally via corrosion of Cu pipes to help control growth of Legionella and other pathogens. However, prior research has shown that Cu does not always reliably control Legionella and sometimes seems to even stimulate its growth. A deeper understanding of the mechanistic effects of Cu on Legionella, at both pure-culture and real-world scales, is critical in order to inform effective controls for Legionella. The overarching objective of the research embodied by this dissertation was aimed at elucidating the chemical and microbial interactions in premise plumbing that govern efficacy of Cu for Legionella control through a series of complementary bench-, pilot-, and field-scale studies. A critical review and synthesis of the literature identified important knowledge gaps in relation to antimicrobial effects of Cu. In particular, changes in the pH, phosphate corrosion control, and rising levels of natural organic matter (NOM) in distributed water are predicted to be important controlling factors. The type of sacrificial anode rod material employed in water heaters was also identified as an underappreciated factor, which directly affects pH, evolution of hydrogen gas as a microbial nutrient, and release of metals (such as aluminum) that bind copper. Microbiological factors: including growth phase of Legionella (e.g., exponential or stationary), strain-specific Cu tolerance, background microbiome composition, and the possibility that viable but non-culturable (VBNC) Legionella might still cause human disease, were also identified as major confounding factors. These knowledge gaps are addressed from various dimensions across each chapter of the dissertation. The effects of pH, orthophosphate corrosion inhibitor concentration, and NOM were examined in bench-scale pure culture experiments over a range of conditions relevant to drinking water. Cupric ions and antimicrobial effects were drastically reduced at pH >7.5, especially in the presence of phosphate, which precipitates copper, or NOM, which complexes the Cu in a form that is less bioavailable. Chick-Watson disinfection models indicated that soluble Cu was the most robust correlate with observed Cu antimicrobial effects across a range of tested waters. This new knowledge suggests that measuring soluble rather than total Cu would be much more informative to guide practitioners in dosing. The research also demonstrated that changes in pH or orthophosphate that have been made to control corrosion over the last few decades, have significantly altered Cu chemistry in buildings, undermining antimicrobial capacity and increasing likelihood of Legionella growth. Pilot-scale experiments confirmed that soluble Cu is an effective indicator of Cu antimicrobial capacity, even in more complex environments represented by realistic hot water plumbing systems. In particular, dosing of orthophosphate, which is widely added by drinking water utilities to control corrosion, directly reduces soluble copper and overall antimicrobial capacity. In some cases, Cu added together with orthophosphate apparently promoted the growth of Legionella, providing an example of at least one circumstance where Cu addition can induce interactive effects that elevate Legionella compared to a control system with trace Cu. It was also demonstrated for the first time that different water heater sacrificial anode types are subject to different corrosion processes, which indirectly influence Cu antimicrobial capacity. Specifically, aluminum ions released from aluminum anode corrosion at 1 mg/L can form an Al(OH)3 gel, which can remove >80% of the soluble Cu from water and reduce Cu antimicrobial effects by >2-log at pH=7. Corrosion from magnesium anodes was found to dramatically increase the pH from 6.8 to >8, which correspondingly reduces Cu antimicrobial capacity. Cu deposition further increased the anode corrosion rate and promoted evolution of hydrogen gas, which is a potent electron donor that stimulates autotrophic microbial growth especially with a magnesium anode. Electric powered anodes did not release metals or alter pH and thus did not diminish Cu antimicrobial capacity. Still, across the pilot-scale experiments, even very high levels of Cu (>1.2 mg/L) at low pH (<7) failed to fully eradicate culturable Legionella. The much lower than expected antimicrobial efficacy of Cu in the pilot-scale hot water plumbing systems was found to be partially explained by the properties of the strain that colonized the systems. Based on fitting the data to a Chick-Watson disinfection model, the outbreak-associated strain that was inoculated into the systems was estimated to be 7 times more tolerant to Cu compared to the common lab strain applied in the bench-scale tests. Further, exponential growth phase L. pneumophila were found to be 2.5 times more susceptible to Cu relative to early stationary phase cultures. It is important to also recognize that, in the pilot-scale systems, drinking water biofilms and the amoeba hosts that colonize them can further shield Legionella from the antimicrobial effects of Cu. Application of shotgun metagenomic sequencing offered the opportunity to more deeply examine the response of Legionella and other pathogens to Cu dosed to the pilot-scale hot water systems in the context of the broader microbiome. It was found that metagenomic analysis provided a sensitive indication of the bioavailability of Cu to the broader microbial community inhabiting the hot water systems, further confirming that the outbreak-associated strain of Legionella that colonized the rigs was relatively tolerant of Cu. Functional gene analysis provided further insight into the mechanistic action of Cu, suggesting multi-modal action of both membrane damage and interruption of nucleic acid replication. The metagenomic analysis further revealed that protozoan host numbers tended to increase in the pilot-scale systems with time, and this could also increase the potential for Legionella proliferation with time. Additional pure culture studies aiming to further assess the mechanistic action of Cu provided strong evidence that Cu can induce a VBNC state for Legionella. This is a concern, given that other studies have indicated that VBNC Legionella are still capable of causing legionellosis. However, VBNC cells are not detected by conventional culturing. Multiple lines of evidence supported the conclusion that Cu induced a VBNC state for Legionella, including membrane integrity, enzyme activity, ATP generation, and Amoebae resuscitation assays applied to two different strains of L. pneumophila. After exposure to Cu, up to a 5-log (99.999%) reduction in culturable Legionella was observed, whereas corresponding reductions in the various viability measures were only by <1-log (90%). In other words, conventional culturing may miss up to 99.99% of the Legionella that is still capable of causing disease. To our knowledge, this is the first study that has assessed the potential for Cu-induced VBNC Legionella. Additional research is needed to further quantify the contribution of VBNC status to challenges in effective Cu-based control of Legionella in premise plumbing. This research further examines, for the first time, the proteomic response of Legionella to Cu, comparing both presumably VBNC and culturable cells. Functional annotation of proteins that were differentially produced by the cells in response to Cu addition revealed that VBNC L. pneumophila modulated its proteome to favor cell membrane- and motility-related proteins, while reducing production of other proteins related to primary metabolism compared to culturable cells. These observations are consistent with the metagenomic-based observations and support the hypothesis that Cu inactivates cells by damaging the cell membrane. The findings also confirmed reduced general cell metabolism that is characteristic of a VBNC state. This dissertation highlights the important and complex effects of Cu on Legionella growth in potable water systems as modified by water chemistry, water heater anode type, characteristics of the surrounding microbiome, and Legionella strain characteristics and growth status. The findings raise important questions about how to measure disinfectant efficacy and provide fundamental new knowledge that can help to better optimize the application of Cu as an antimicrobial to drinking water systems and better protect public health. / Doctor of Philosophy / The opportunistic pathogen Legionella is the leading cause of reportable waterborne disease outbreaks in the United States. Legionella is capable of growing in drinking water plumbing systems in homes, hospitals, hotels, and other buildings. Legionella is spread by inhaling tiny droplets of water that are suspended in the air when using the water, for example when showering, resulting in a severe and deadly form of pneumonia called Legionnaires' Disease. Copper is a promising antimicrobial that can be dosed directly into a building's water system by installing a copper-silver ionization system. There is also interest in understanding whether copper released naturally from copper pipes could help control Legionella. However, prior research indicates that copper sometimes inhibits, sometimes has no effect, and sometimes even seems to stimulate Legionella growth. The purpose of this dissertation was to better understand how the chemical properties of the drinking water, such as pH, presence of corrosion inhibitors that are commonly added to the water by utilities, and natural organic matter impact the ability of copper to kill Legionella. Impacts of the design of the drinking water system were also examined, for example, the material used in the anodes of water heaters to prevent corrosive damage to other system components was hypothesized to change the water chemistry in such a way that could also interfere with copper disinfection. Finally, the effect of the strain of Legionella, its growth phase (exponential or stationary), and culturability status (culturable versus viable but non-culturable) was also examined. Experiments were conducted over a wide range of conditions, from bench-scale pure culture experiments of a few days to full-scale plumbing systems over a period of 3.5 years. The complementary approaches maximize the strength of scientific conclusions about approaches that can more effectively control Legionella. Several discoveries were made as a result of this research that can help to improve the use of copper for controlling Legionella in drinking water systems. In particular, it was found that it is best to keep the pH less than 7.5, because above pH 7.5 copper reacts with orthophosphate corrosion inhibitor or natural organic matter in the water in a manner that makes it less potent to microbes. Through disinfection modeling it was found that soluble copper was the best predictor of the ability to kill Legionella. Therefore, it is recommended from this research that practitioners should monitor soluble copper instead of total copper for the purpose of assessing Legionella control. From the pilot-scale experiments, it was further found that the type of anode installed in the water heater can affect the ability of copper to kill Legionella. Magnesium anodes performed the worst, likely because they raised the pH above the recommended level of 7.5. Aluminum anodes were also a problem because aluminum ions released form an aluminum hydroxide gel that can remove more than 80% of the soluble copper from water. Electric powered anodes did not reduce copper antimicrobial effects by raising pH or forming a gel, but they are much less commonly used. A surprising finding throughout this study was that very high levels of copper (>1.2 mg/L) were required to measurably reduce Legionella in the pilot-scale systems. In the pure culture experiments, it was found that the outbreak-associated strain from Quincy, IL, that was inoculated into the system was highly copper tolerant. This demonstrated that the strain of Legionella that colonizes a particular drinking water system could be the reason why copper is sometimes less effective. Pure culture experiments also found that stationary phase Legionella are more difficult to kill than exponential phase Legionella, which could explain some discrepancies among lab studies reported in the literature. A particularly noteworthy discovery of this research was that copper can make it appear as if Legionella have been killed, because the traditional culture media indicate that there is no growth on the Petri dish; however, they are in fact still alive and capable of causing human disease. This is referred to as a "viable but non-culturable (VBNC)" state. The VBNC state of Legionella was confirmed using an array of techniques (membrane integrity, enzyme activity, ATP generation, and amoebae resuscitation) for two strains of L. pneumophila. We also examined how VBNC Legionella cellular functions were impacted by copper using whole cell proteome, i.e., analysis of all of the proteins extracted from Legionella. Copper induced VBNC Legionella modulated its proteome to favor cell membrane and motility related proteins, and reduced others related to primary metabolism compared with culturable cells. These results were consistent with those obtained via shotgun metagenomic analysis of the microbial community DNA in the pilot-scale water systems. Given the potential for VBNC organisms to prevail in systems disinfected with copper, it is recommended to supplement culture-based monitoring with molecular-based monitoring, e.g., with quantitative polymerase chain reaction. This dissertation highlights the important and complex effects of copper on Legionella growth in potable water systems. The findings help to inform guidance on how to improve the antimicrobial effect of copper, through adjusting the water chemistry, selecting appropriate water heater anodes, and optimizing the overall hot water system design. The dissertation also helps to inform improved strategies for monitoring the efficacy of copper for killing Legionella in real-world systems. Overall, the findings can help to improve policy and practice aimed at reducing the incidence of Legionnaires' Disease and protecting public health.

Ce travail de thèse concerne l'étude de l'écologie microbienne d'un atelier de découpe de viande bovine, dans le but de mieux comprendre la persistance bactérienne, c'est-à-dire, la présence répétée d'un même clone bactérien pendant une longue période malgré l'application bien conduite et régulière du nettoyage et de la désinfection (N-D). Des prélèvements par « chiffonnages » multiples de surfaces d'équipements ont été réalisés lors de trois campagnes de prélèvement espacées les unes des autres d'au moins six mois. Les prélèvements ont été réalisés sur un tapis convoyeur en polychlorure de vinyle (PVC) et sur des machines éplucheuses en acier inoxydable avant et après N-D. Nous avons quantifié les cellules totales (les cellules vivantes et les cellules mortes) par PCR quantitative en temps réel (qPCR), les cellules viables par EMA-qPCR, et les UFC (provenant de cellules cultivables) par dénombrement après incubation à 25°C sur gélose tryptone soja. Les résultats montrent qu'avant N-D, les cellules totales (en moyenne 5,6 – exprimé en log10 cellules/cm2 – sur PVC et 4,7 sur acier inoxydable) sont plus nombreuses que les cellules viables (4,5 sur PVC et 4,4 sur acier inoxydable) lesquelles sont plus nombreuses que les UFC (3,8 sur PVC et 2,9 sur acier inoxydable). Le N-D entraîne moins d'une réduction décimale (RD) des populations à l'exception des UFC sur acier inoxydable qui subissent 1,5 RD en moyenne. Ce dernier chiffre s'explique par des forces d'adhésion faibles. L'étude de la diversité des bactéries cultivables montre que sur un total de 51 genres identifiés, 13 seulement sont retrouvés lors des trois campagnes de prélèvements. Les isolats de ces 13 genres représentent 75, 72 et 62% des isolats des campagnes1, 2 et 3 respectivement. Parmi ces isolats, les plus fréquents sont (par ordre décroissant du nombre d'isolats) : Pseudomonas, Staphylococcus, Microbacterium, Acinetobacter, Chryseobacterium, Psychrobacter et Kocuria. Le génotypage d'isolats de 3 genres majoritaires (Staphylococcus, Pseudomonas et Acinetobacter) montre qu'une seule souche, Staphylococcus equorum, est sans aucun doute persistante. L'ensemble de ces observations montrent que l'écosystème varie d'une campagne à une autre. Ces modifications de la diversité bactérienne reflèteraient les modifications de flores des viandes traitées dans l'atelier, qui ont des origines multiples. En outre, il apparaît que, contrairement à ce qui est généralement admis, les bactéries à coloration de Gram négative cultivables sont plus facilement inactivées par le N-D que les bactéries à coloration de Gram positive. L'étude de l'écosystème par PCR-DGGE a permis d'identifier sept genres bactériens et montre que les espèces dominantes sont toutes sous forme vivante, autrement dit, aucune des espèces dominantes n'a été détectée uniquement sous forme de cellules mortes. Sur les sept genres identifiés six sont des Gram – dont majoritairement les genres Acinetobacter, Pseudomonas et Psychrobacter. Cette dominance montre que le N-D permet une forte perte de cultivabilité des bactéries Gram – mais qu'une grande partie n'est pas détachée. La dominance des bactéries Gram – observée par PCR-DGGE masque les staphylocoques qui ne sont pas détectés alors qu'ils sont majoritaires parmi la flore cultivable. Seul un genre bactérien, Propionibacterium, est identifié par PCR-DGGE uniquement mais il n'est trouvé qu'à une seule campagne et uniquement sur l'acier inoxydable avant N-D. En conclusion, l'avancée majeure de ce travail est la mise en évidence qu'une proportion importante de bactéries survit après les opérations très poussées de N-D mais pour une période transitoire. / The aim of this work is to acquire a better knowledge of the microbial ecology of a beef processing plant to understand bacterial persistence, e.g. the presence of a clone isolated several times on several visits in the same processing plant despite regular Cleaning and disinfection (C&D) procedures. Successive swabbing were performed on a PVC conveyor belt and skinning machines made of stainless steel before and after C&D during three surveys in minimal 6 month-intervals. Total cells (live and dead cells) were quantified using real-time quantitative PCR (qPCR). Viable cells e.g. cells with intact membrane, were assessed using Ethidium Monoazide combined with qPCR. Culturable cells (CFU) were determined from plate counts on Tryptone Soy Agar. Before C&D, total cells (5.6 log cells/cm2 and 4.7 log10 cells/cm2 on PVC and stainless steel respectively) were greater than viable cells (4.5 and 4.4 log10 cells/cm2) and CFUs (3.8 and 2.9 log10 CFU/cm2). C&D lead to less than 1 log10 reduction in bacterial populations except for CFU counts on stainless steel where a 1.5 log reduction is observed. This result is highlighted by the weak attachment strengths observed on stainless steel for CFUs. Identification of the culturable microbiota revealed that out of 51 genera identified, 13 were found at all the visits. These genera represented 75, 72 and 62% of the total isolates. The most frequently identified bacteria were Pseudomonas, Staphylococcus, Microbacterium, Acinetobacter, Chryseobacterium, Psychrobacter and Kocuria. Molecular typing of three dominant genera (Staphylococcus, Pseudomonas and Acinetobacter) showed that only one strain, Staphylococcus equorum, was persistent in the premises. Our results show that the microbial ecosystem is different from one survey to another, which reflect the various geographical origins of meat products. Contrary to widespread belief, Gram negative strains were more easily eliminated by C&D than Gram positive strains. Furthermore, the microbial diversity assessed by PCR-DGGE allowed the identification of 7 genera. This molecular approach showed that dominant species are all in a viable state: none of these species was solely detected in a dead state. Of the 7 genera identified, 6 were Gram negative, Acinetobacter, Pseudomonas and Psychrobacter being predominant. This result highlights that C&D induced the lost of culturability of Gram negative bacteria although a high proportion was not detached from the surface. The predominance of Gram negative microflora, didn’t allow the detection of staphylococcal isolates which were numerous in the culturable microflora. One genus, Propionibacterium, isolated in one survey on stainless steel before C&D was only identified by PCR-DGGE. In conclusion, the present study has demonstrated that a large proportion of bacteria can survive drastic cleaning and disinfection for a transient period.

O desenvolvimento municipal sustentável é um fenômeno complexo. Para que seja possível entender tal fenômeno, é necessário levar em consideração como se comportam as diversas variáveis de dimensões distintas e a interação de uma grande quantidade de atores envolvidos em um processo dinâmico, transformando constantemente o sistema social de uma determinada região. Esta dissertação tem em sua essência, ser um trabalho que está de acordo com o novo paradigma da ciência, descartando explicações simples e prontas sobre o desenvolvimento. Busca agregar toda a complexidade, instabilidade e intersubjetividade que o assunto compreende. Assim, o principal objetivo deste trabalho é desenvolver e diagnosticar o sistema de administração das prefeituras municipais para o desenvolvimento sustentável, tendo como base os objetivos de desenvolvimento sustentável (ODS), utilizando como métodos o Viable System Model (VSM) em prefeituras municipais e a criação de Indicador Sistêmico-Cibernético de Desenvolvimento Sustentável (ISCDS) por meio da análise fatorial (AF) para os municípios do estado de São Paulo. Os objetivos específicos também são desenvolvidos de maneira a contribuir ao objetivo principal. Dentre eles estão: diferenciar os diversos conceitos de desenvolvimento, identificar e delimitar o sistema onde acontece o desenvolvimento local, verificar como os ODS podem ser uma forma de promover o desenvolvimento sistêmico, identificar as principais variáveis secundárias que podem compor o desenvolvimento local e que estejam ligadas a cada um dos objetivos de desenvolvimento sustentável da ONU, desenvolver um indicador capaz de medir o desenvolvimento sistêmico utilizando os ODS e, por fim, identificar as diferenças entre os municípios do estado de São Paulo criando agrupamentos de municípios semelhantes. Para tanto, este é um trabalho sistêmico tanto em sua filosofia quanto estrutura, adotando métodos e técnicas tanto quantitativas como qualitativas para se chegar aos resultados das perguntas de pesquisa. Para que fosse possível obter conhecimento suficiente acerca dos temas tratados e propor uma base metodológica sólida de modo a mostrar os resultados esperados, optou-se por realizar um amplo referencial teórico, envolvendo três itens: desenvolvimento, abordagens sistêmico-cibernéticas e objetivos de desenvolvimento sustentável. Os resultados mostraram que é possível aplicar o VSM no sistema administrativo das prefeituras municipais, sendo uma poderosa ferramenta para administrar o a prefeitura de maneira eficiente e também útil para identificar diversas falhas sistêmicas, conforme são expostas nas considerações finais deste trabalho. Por meio da análise fatorial, tendo como base os ODS, foi possível desenvolver um modelo para se medir o desenvolvimento sustentável sistêmico municipal: o ISCDS para os 645 municípios do estado de São Paulo. A partir da obtenção do ISCDS, foram encontrados sete clusters com características diferentes, indicando a necessidade de tratativas administrativas diferentes que visem o desenvolvimento sustentável para cada agrupamento encontrado. / Sustainable municipal development is a complex phenomenon. To understand this phenomenon, it is necessary to take into account not only the way of huge number variables of different dimensions behave, but also how the large number of actors are involved in a dynamic process, constantly transforming the social system of a given region. This dissertation has in its essence, to be a work that is in accordance with the new paradigm of science, discarding simple and ready explanations about development. It seeks to add all the complexity, instability and intersubjectivity that the subject comprehends. Thus, the main objective of this work is to develop and diagnose the municipal town hall administration system for the sustainable development, based on the sustainable development goals (SDG). The methods used are the Viable System Model (VSM) for the municipal town hall management, and the factor analysis to model the Systemic-Cybernetic Indicator of Sustainable Development (SCISD) for the municipalities of the state of São Paulo. Specific objectives are also developed in order to contribute to the main objective. These include: a) to differentiate the various development concepts, b) to identify the system where local development takes place, b) to check how the SDGs can promote systemic development, c) to identify the main variables that could be used to measure sustainable development, and which of them are connected to each of the UN\'s SDGs and, finally, d) to identify the differences between the municipalities of the state of São Paulo, creating clusters of similar municipalities according to the SCISD developed. To do so, this is a systemic work both in its philosophy and structure. It was adopted methods and techniques both quantitative and qualitative to explore to the results of the research questions. In order to obtain sufficient knowledge about the topics addressed and to propose a solid methodological basis to show the expected results, it was decided to make a broad theoretical reference, involving three items: development, systems thinking and cybernetics approaches and sustainable development goals. The results showed that it is possible to apply the VSM in the administrative system of the city halls, being a powerful tool not only to manage the city in an efficient way, but also useful to identify several systemic failures, as they are exposed in the final considerations of this work. Through the factorial analysis, based on the SDGs, it was also possible to develop a model to measure the level of municipal sustainable development: the SCISD for the 645 municipalities of the state of São Paulo. Also, from the SCISD, seven clusters with different characteristics were found, indicating the need for different administrative approaches according to the different at sustainable development characteristics found in each cluster.

Based on literature, a theoretical model was developed for viable performance consisting of eight constructs whilst the trust model of Martins (2000) was used to measure four organisational trust constructs. Exploratory factor analysis was used to extract the constructs, and structural equation modelling was employed to validate the models against the data. An empirical model for viable performance resulted in a solution with seven constructs and organisational trust with five constructs. The two empirical models were unified into a model of viable performance and trust resulting in a measurement model where all 12 constructs were shown. Significant levels of internal consistency were measured. The resulting measurement model was tested for group differences, and no significant differences were found, indicating that the assessment can be used across different groups. It was concluded that the aim to construct and test an integrated and comprehensive theoretical framework of viable performance and trust was achieved and the resulting Viable Performances and Trust Indicator (VPTI) was validated as an assessment to be used across groups. Organisations can thus use the framework and VPTI assessment tool with confidence to assess performance and trust across different biographical groups. Future researchers can build on this exploratory study to refine the scales and apply the measurement model within the wider context of South Africa or as a globally accepted model. / IndustriaI and Organisational Psychology / D. Litt. et Phil. (IndustriaI and Organisational Psychology)

O desenvolvimento municipal sustentável é um fenômeno complexo. Para que seja possível entender tal fenômeno, é necessário levar em consideração como se comportam as diversas variáveis de dimensões distintas e a interação de uma grande quantidade de atores envolvidos em um processo dinâmico, transformando constantemente o sistema social de uma determinada região. Esta dissertação tem em sua essência, ser um trabalho que está de acordo com o novo paradigma da ciência, descartando explicações simples e prontas sobre o desenvolvimento. Busca agregar toda a complexidade, instabilidade e intersubjetividade que o assunto compreende. Assim, o principal objetivo deste trabalho é desenvolver e diagnosticar o sistema de administração das prefeituras municipais para o desenvolvimento sustentável, tendo como base os objetivos de desenvolvimento sustentável (ODS), utilizando como métodos o Viable System Model (VSM) em prefeituras municipais e a criação de Indicador Sistêmico-Cibernético de Desenvolvimento Sustentável (ISCDS) por meio da análise fatorial (AF) para os municípios do estado de São Paulo. Os objetivos específicos também são desenvolvidos de maneira a contribuir ao objetivo principal. Dentre eles estão: diferenciar os diversos conceitos de desenvolvimento, identificar e delimitar o sistema onde acontece o desenvolvimento local, verificar como os ODS podem ser uma forma de promover o desenvolvimento sistêmico, identificar as principais variáveis secundárias que podem compor o desenvolvimento local e que estejam ligadas a cada um dos objetivos de desenvolvimento sustentável da ONU, desenvolver um indicador capaz de medir o desenvolvimento sistêmico utilizando os ODS e, por fim, identificar as diferenças entre os municípios do estado de São Paulo criando agrupamentos de municípios semelhantes. Para tanto, este é um trabalho sistêmico tanto em sua filosofia quanto estrutura, adotando métodos e técnicas tanto quantitativas como qualitativas para se chegar aos resultados das perguntas de pesquisa. Para que fosse possível obter conhecimento suficiente acerca dos temas tratados e propor uma base metodológica sólida de modo a mostrar os resultados esperados, optou-se por realizar um amplo referencial teórico, envolvendo três itens: desenvolvimento, abordagens sistêmico-cibernéticas e objetivos de desenvolvimento sustentável. Os resultados mostraram que é possível aplicar o VSM no sistema administrativo das prefeituras municipais, sendo uma poderosa ferramenta para administrar o a prefeitura de maneira eficiente e também útil para identificar diversas falhas sistêmicas, conforme são expostas nas considerações finais deste trabalho. Por meio da análise fatorial, tendo como base os ODS, foi possível desenvolver um modelo para se medir o desenvolvimento sustentável sistêmico municipal: o ISCDS para os 645 municípios do estado de São Paulo. A partir da obtenção do ISCDS, foram encontrados sete clusters com características diferentes, indicando a necessidade de tratativas administrativas diferentes que visem o desenvolvimento sustentável para cada agrupamento encontrado. / Sustainable municipal development is a complex phenomenon. To understand this phenomenon, it is necessary to take into account not only the way of huge number variables of different dimensions behave, but also how the large number of actors are involved in a dynamic process, constantly transforming the social system of a given region. This dissertation has in its essence, to be a work that is in accordance with the new paradigm of science, discarding simple and ready explanations about development. It seeks to add all the complexity, instability and intersubjectivity that the subject comprehends. Thus, the main objective of this work is to develop and diagnose the municipal town hall administration system for the sustainable development, based on the sustainable development goals (SDG). The methods used are the Viable System Model (VSM) for the municipal town hall management, and the factor analysis to model the Systemic-Cybernetic Indicator of Sustainable Development (SCISD) for the municipalities of the state of São Paulo. Specific objectives are also developed in order to contribute to the main objective. These include: a) to differentiate the various development concepts, b) to identify the system where local development takes place, b) to check how the SDGs can promote systemic development, c) to identify the main variables that could be used to measure sustainable development, and which of them are connected to each of the UN\'s SDGs and, finally, d) to identify the differences between the municipalities of the state of São Paulo, creating clusters of similar municipalities according to the SCISD developed. To do so, this is a systemic work both in its philosophy and structure. It was adopted methods and techniques both quantitative and qualitative to explore to the results of the research questions. In order to obtain sufficient knowledge about the topics addressed and to propose a solid methodological basis to show the expected results, it was decided to make a broad theoretical reference, involving three items: development, systems thinking and cybernetics approaches and sustainable development goals. The results showed that it is possible to apply the VSM in the administrative system of the city halls, being a powerful tool not only to manage the city in an efficient way, but also useful to identify several systemic failures, as they are exposed in the final considerations of this work. Through the factorial analysis, based on the SDGs, it was also possible to develop a model to measure the level of municipal sustainable development: the SCISD for the 645 municipalities of the state of São Paulo. Also, from the SCISD, seven clusters with different characteristics were found, indicating the need for different administrative approaches according to the different at sustainable development characteristics found in each cluster.

Les nouvelles pratiques utilisées pour l’élaboration du vin amènent à une recrudescence des altérations microbiennes. C’est pourquoi, de nouvelles méthodes doivent être développées afin de quantifier ces microorganismes de façon précise, rapide et avec de faibles coûts. Les principales altérations du vin sont dues aux bactéries acétiques (BA) (A. aceti, A. pasteurianus, G. oxydans et Ga. liquefaciens) et à Brettanomyces bruxellensis. Par l’action d’enzymes, les 1ères transforment l’éthanol en acide acétique alors que B. bruxellensis transforme les acides hydroxycinnamiques en éthyles phénols (EP) (molécules odorantes désagréables). La cytométrie en flux couplée à la technique d’hybridation in situ en fluorescence a tout d’abord été étudiée. Aucun résultat reproductible n’a été développé pour les BA en vin rouge alors que pour B. bruxellensis, le protocole existant a été amélioré avec une quantification possible en 18 h. La PCR en temps réel a également été utilisées afin de quantifier ces microorganismes. Un protocole a été développé pour la quantification des BA en vin rouge (103 cellules/mL) avec l’utilisation d’un témoin interne microbiologique permettant de valider le rendement de l’extraction de l’ADN. Pour B. bruxellensis, trois kits commerciaux ont été analysés lors d’une étude interlaboratoires. Les quantifications se sont révélées significativement différentes des énumérations sur boite de Pétri avec une quantification des cellules mortes. De plus, il a été étudié et validé l’effet population de B. bruxellensis sur l’efficacité du SO2. Il ressort également de ces expérimentations que les cellules en état viable mais non cultivable ne produisent pas d’EP. / New practices used to elaborate wine lead to an increase of wine spoilage due to microorganisms. That is why, new technics have to be developed to quantify these microorganisms accurately, quickly and with low costs. The main wine spoilages are due to acetic acid bacteria (AAB) (A. aceti, A. pasteurianus, G. oxydans and Ga. liquefaciens) and Brettanomyces bruxellensis development. AAB transforms ethanol to acetic acid while B. bruxellensis transforms hydroxycinnamic acids to ethyl phenols (EP) (unpleasant odor molecules). In order to detect these wine spoilage microrganisms, flow cytometry coupled to fluorescent in situ hybridization has been assessed. No reproducible results have been developed for AAB in red wine while for B. bruxellensis, the existing protocol has been improved with a possible quantification after 18 h compared to 48-72 h in the previous protocol. The real-time PCR was also used to quantify these microorganisms. A protocol has been developed for the AAB quantification in red wine (103 cells/mL) with the use of a microbiological internal control to validate the DNA yield after extraction. For B. bruxellensis, three commercial kits were analyzed in an interlaboratory study. Quantifications were significantly different to the enumerations by Petri dish, with dead cell quantifications. Moreover, we demonstrated that the effectiveness of sulfite is dependent of the B. bruxellensis population. It also appears from these experiments that cells in viable but not culturable state do not produce EP.

The aim of this thesis is to monitor how the amount of water (water activity) affects the amount of microorganisms is selected packaged meat products from the point of view standards, which specify their specific requirements for the health safety of these products. The first part of this work describes various types of packaging materials of meat products, the microorganisms that occur in them and the definition of water activity. The second part describes the actual measurement of water activity and the measurement of the occurrence of microorganisms and coliforms, from selection of suitable meat products, their types of packaging, sample preparation and evaluation of measured methods. The results showed, that the activity of water affects the occurrence of microorganisms and coliforms, in that the higher the activity of water, the higher microorganisms presence.

A presente obra aborda a experiência recente de empresas brasileiras no gerenciamento continuado de ambientes multiprojetos pelo Método da Corrente Crítica, ou CCPM, Critical Chain Path Method, através de um estudo de casos múltiplos, conduzido frente às empresas EMBRAER e EMBRACO. O desenvolvimento do estudo bibliográfico permitiu pontuar as diferenças conceituais de gerenciamento de portfólios, gerenciamento de programas, gerenciamento de projetos e gerenciamento de ambientes multiprojetos, posicionar o CCPM dentro do rol de algoritmos de solução de ambientes multiprojetos e identificar as características que expliquem sua maior difusão no universo da administração de projetos, em detrimento de outras proposições matematicamente até mais robustas que ele. A pesquisa de campo identificou que, dentre as seis práticas preconizadas pela filosofia gerencial Visão Viável para a condução do CCPM, três delas despontam como fatores críticos de sucesso para o gerenciamento de ambientes multiprojetos: reduzindo a multitarefa nociva, planejando e executando. Por fim, os dados coletados evidenciaram que o CCPM traz ganhos quantitativamente significativos ao desempenho do gerenciamento de ambientes multiprojetos, especialmente no que diga respeito ao incremento da capacidade de entrega de projetos (throughput) de um pool de recursos de tamanho constante, e que sua efetiva implementação é impactada pela estrutura organizacional e pela maturidade em administração de projetos da empresa. / The present work discusses the recent experience of Brazilian companies in continued management of multi-project environments by means of Critical Chain Path Method, or CCPM, through a multiple case study conducted on the companies EMBRAER and EMBRACO. The bibliographic study allowed the determination of conceptual differences among portfolio management, program management, project management and multi-project environment management, positioned the CCPM inside the roll of multi-project solution algorithms and identified characteristics that explain its widest adoption in the universe of project management, to the detriment of other propositions, although mathematically even more robust than CCPM itself. The field research identified that, among the six practices advocated by the managerial philosophy Viable Vision for the conduction of CCPM, three of them stand out as critical success factors for managing multi-project environments: reducing harmful multitasking, planning and executing. Finally, the data collected indicated that the CCPM brings significant performance gains to the management of multi-project environments, especially related to the throughput increase of project delivery, considering a constant size pool of resources, and that their effective implementation is impacted by the organizational structure and the maturity in project management of the company.

A presente obra aborda a experiência recente de empresas brasileiras no gerenciamento continuado de ambientes multiprojetos pelo Método da Corrente Crítica, ou CCPM, Critical Chain Path Method, através de um estudo de casos múltiplos, conduzido frente às empresas EMBRAER e EMBRACO. O desenvolvimento do estudo bibliográfico permitiu pontuar as diferenças conceituais de gerenciamento de portfólios, gerenciamento de programas, gerenciamento de projetos e gerenciamento de ambientes multiprojetos, posicionar o CCPM dentro do rol de algoritmos de solução de ambientes multiprojetos e identificar as características que expliquem sua maior difusão no universo da administração de projetos, em detrimento de outras proposições matematicamente até mais robustas que ele. A pesquisa de campo identificou que, dentre as seis práticas preconizadas pela filosofia gerencial Visão Viável para a condução do CCPM, três delas despontam como fatores críticos de sucesso para o gerenciamento de ambientes multiprojetos: reduzindo a multitarefa nociva, planejando e executando. Por fim, os dados coletados evidenciaram que o CCPM traz ganhos quantitativamente significativos ao desempenho do gerenciamento de ambientes multiprojetos, especialmente no que diga respeito ao incremento da capacidade de entrega de projetos (throughput) de um pool de recursos de tamanho constante, e que sua efetiva implementação é impactada pela estrutura organizacional e pela maturidade em administração de projetos da empresa. / The present work discusses the recent experience of Brazilian companies in continued management of multi-project environments by means of Critical Chain Path Method, or CCPM, through a multiple case study conducted on the companies EMBRAER and EMBRACO. The bibliographic study allowed the determination of conceptual differences among portfolio management, program management, project management and multi-project environment management, positioned the CCPM inside the roll of multi-project solution algorithms and identified characteristics that explain its widest adoption in the universe of project management, to the detriment of other propositions, although mathematically even more robust than CCPM itself. The field research identified that, among the six practices advocated by the managerial philosophy Viable Vision for the conduction of CCPM, three of them stand out as critical success factors for managing multi-project environments: reducing harmful multitasking, planning and executing. Finally, the data collected indicated that the CCPM brings significant performance gains to the management of multi-project environments, especially related to the throughput increase of project delivery, considering a constant size pool of resources, and that their effective implementation is impacted by the organizational structure and the maturity in project management of the company.